4.1.3. Buffer solutions EUROPEAN PHARMACOPOEIA 5.

Vanadium standard solution (1 g/l V). 5003300. Buffer solution pH 2.5 R1. 4000400.
Dissolve in water R ammonium vanadate R equivalent to To 4.9 g of dilute phosphoric acid R add 250 ml of water R.
0.230 g of NH4VO3 and dilute to 100.0 ml with the same Adjust the pH (2.2.3) with dilute sodium hydroxide
solvent. solution R and dilute to 500.0 ml with water R.
Zinc standard solution (5 mg/ml Zn). 5003400. Phosphate buffer solution pH 2.8. 4010600.
Dissolve 3.15 g of zinc oxide R in 15 ml of hydrochloric Dissolve 7.8 g of sodium dihydrogen phosphate R in 900 ml
acid R and dilute to 500.0 ml with water R. of water R, adjust to pH 2.8 (2.2.3) with phosphoric acid R
and dilute to 1000 ml with the same solvent.
Zinc standard solution (100 ppm Zn). 5003401.
Immediately before use, dilute with water R to 10 times its Buffer solution pH 3.0. 4008000.
volume a solution containing zinc sulphate R equivalent to Dissolve 21.0 g of citric acid R in 200 ml of 1 M sodium
0.440 g of ZnSO4,7H2O and 1 ml of acetic acid R in 100.0 ml. hydroxide and dilute to 1000 ml with water R. Dilute 40.3 ml
of this solution to 100.0 ml with 0.1 M hydrochloric acid.
Zinc standard solution (10 ppm Zn). 5003402.
Immediately before use, dilute zinc standard solution 0.25 M Citrate buffer solution pH 3.0. 4012600.
(100 ppm Zn) R to 10 times its volume with water R. Dissolve 4.8 g of citric acid R in 80 ml of water R. Adjust
Zinc standard solution (5 ppm Zn). 5003403. the pH (2.2.3) with 1 M sodium hydroxide and dilute to
100.0 ml with water R.
Immediately before use, dilute zinc standard solution
(100 ppm Zn) R to 20 times its volume with water R. 0.1 M Phosphate buffer solution pH 3.0. 4011500.
Zirconium standard solution (1 g/l Zr). 5003500. Dissolve 12.0 g of anhydrous sodium dihydrogen
phosphate R in water R, adjust the pH (2.2.3) with dilute
Dissolve zirconyl nitrate R equivalent to 0.293 g of phosphoric acid R1 and dilute to 1000 ml with water R.
ZrO(NO3)2,2H2O in a mixture of 2 volumes of hydrochloric
acid R and 8 volumes of water R and dilute to 100.0 ml with Phosphate buffer solution pH 3.0. 4000500.
the same mixture of solvents.
Mix 0.7 ml of phosphoric acid R with 100 ml of water R.
Dilute to 900 ml with the same solvent. Adjust to pH 3.0
(2.2.3) with strong sodium hydroxide solution R and dilute
04/2007:40103 to 1000 ml with water R.
Phosphate buffer solution pH 3.0 R1. 4010000.
4.1.3. BUFFER SOLUTIONS Dissolve 3.40 g of potassium dihydrogen phosphate R in
900 ml of water R. Adjust to pH 3.0 (2.2.3) with phosphoric
Buffered acetone solution. 4000100. acid R and dilute to 1000.0 ml with water R.
Dissolve 8.15 g of sodium acetate R and 42 g of sodium
chloride R in water R, add 68 ml of 0.1 M hydrochloric acid Phosphate buffer solution pH 3.2. 4008100.
and 150 ml of acetone R and dilute to 500 ml with water R. To 900 ml of a 4 g/l solution of sodium dihydrogen
Buffer solution pH 2.0. 4000200. phosphate R, add 100 ml of a 2.5 g/l solution of phosphoric
acid R. Adjust the pH (2.2.3) if necessary.
Dissolve 6.57 g of potassium chloride R in water R and add
119.0 ml of 0.1 M hydrochloric acid. Dilute to 1000.0 ml Phosphate buffer solution pH 3.2 R1. 4008500.
with water R.
Adjust a 35.8 g/l solution of disodium hydrogen
Phosphate buffer solution pH 2.0. 4007900. phosphate R to pH 3.2 (2.2.3) with dilute phosphoric acid R.
Dilute 100.0 ml of the solution to 2000.0 ml with water R.
Dissolve 8.95 g of disodium hydrogen phosphate R and
3.40 g of potassium dihydrogen phosphate R in water R Buffer solution pH 3.5. 4000600.
and dilute to 1000.0 ml with the same solvent. If necessary
adjust the pH (2.2.3) with phosphoric acid R. Dissolve 25.0 g of ammonium acetate R in 25 ml of water R
and add 38.0 ml of hydrochloric acid R1. Adjust the pH
Sulphate buffer solution pH 2.0. 4008900. (2.2.3) if necessary with dilute hydrochloric acid R or dilute
ammonia R1. Dilute to 100.0 ml with water R.
Dissolve 132.1 g of ammonium sulphate R in water R
and dilute to 500.0 ml with the same solvent (Solution I). Phosphate buffer solution pH 3.5. 4000700.
Carefully and with constant cooling stir 14 ml of sulphuric
acid R into about 400 ml of water R ; allow to cool and dilute Dissolve 68.0 g of potassium dihydrogen phosphate R in
to 500.0 ml with water R (Solution II). Mix equal volumes of water R and dilute to 1000.0 ml with the same solvent.
solutions I and II. Adjust the pH (2.2.3) if necessary. Adjust the pH (2.2.3) with phosphoric acid R.

Buffer solution pH 2.2. 4010500. Buffer solution pH 3.6. 4000800.

Mix of 6.7 ml of phosphoric acid R with 50.0 ml of a 4 per To 250.0 ml of 0.2 M potassium hydrogen phthalate R add
cent solution of dilute sodium hydroxide solution R and 11.94 ml of 0.2 M hydrochloric acid. Dilute to 1000.0 ml
dilute to 1000.0 ml with water R. with water R.

Buffer solution pH 2.5. 4000300. Buffer solution pH 3.7. 4000900.

Dissolve 100 g of potassium dihydrogen phosphate R in To 15.0 ml of acetic acid R add 60 ml of alcohol R and
800 ml of water R ; adjust to pH 2.5 (2.2.3) with hydrochloric 20 ml of water R. Adjust to pH 3.7 (2.2.3) by the addition of
acid R and dilute to 1000.0 ml with water R. ammonia R. Dilute to 100.0 ml with water R.

4932 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 5.7
Buffered copper sulphate solution pH 4.0. 4001000.
Dissolve 0.25 g of copper sulphate R and 4.5 g of ammonium
acetate R in dilute acetic acid R and dilute to 100.0 ml with
the same solvent.
Acetate buffer solution pH 4.4. 4001100.
Dissolve 136 g of sodium acetate R and 77 g of ammonium
acetate R in water R and dilute to 1000.0 ml with the same
solvent ; add 250.0 ml of glacial acetic acid R and mix.
Phthalate buffer solution pH 4.4. 4001200.
Dissolve 2.042 g of potassium hydrogen phthalate R in
50 ml of water R, add 7.5 ml of 0.2 M sodium hydroxide and Dissolve 54.4 g of sodium acetate R in 50 ml of water R,
dilute to 200.0 ml with water R.
Acetate buffer solution pH 4.5. 4012500.
Dissolve 77.1 g of ammonium acetate R in water R. Add
70 ml of glacial acetic acid R and dilute to 1000.0 ml with Solution I. Dissolve 13.61 g of potassium dihydrogen
water R.
0.05 M Phosphate buffer solution pH 4.5. 4009000.
Dissolve 6.80 g of potassium dihydrogen phosphate R in
1000.0 ml of water R. The pH (2.2.3) of the solution is 4.5.
Sodium acetate buffer solution pH 4.5. 4010100.
Dissolve 63 g of anhydrous sodium acetate R in water R,
add 90 ml acetic acid R and adjust to pH 4.5, and dilute to
1000 ml with water R.
Acetate buffer solution pH 4.6. 4001400.
Dissolve 5.4 g of sodium acetate R in 50 ml of water R, add
2.4 g of glacial acetic acid R and dilute to 100.0 ml with
water R. Adjust the pH (2.2.3) if necessary.
Succinate buffer solution pH 4.6. 4001500.
Disssolve 11.8 g of succinic acid R in a mixture of 600 ml
of water R and 82 ml of 1 M sodium hydroxide and dilute
to 1000.0 ml with water R.
Acetate buffer solution pH 4.7. 4001600.
Dissolve 136.1 g of sodium acetate R in 500 ml of water R.
Mix 250 ml of this solution with 250 ml of dilute acetic
acid R. Shake twice with a freshly prepared, filtered, 0.1 g/l
solution of dithizone R in chloroform R. Shake with carbon
tetrachloride R until the extract is colourless. Filter the
aqueous layer to remove traces of carbon tetrachloride.
Acetate buffer solution pH 5.0. 4009100.
To 120 ml of a 6 g/l solution of glacial acetic acid R add
100 ml of 0.1 M potassium hydroxide and about 250 ml of
water R. Mix. Adjust the pH to 5.0 with a 6 g/l solution of
acetic acid R or with 0.1 M potassium hydroxide and dilute
to 1000.0 ml with water R.
Citrate buffer solution pH 5.0. 4010700.
Prepare a solution containing 20.1 g/l of citric acid R and
8.0 g/l of sodium hydroxide R. Adjust the pH with dilute
hydrochloric acid R.
Phosphate buffer solution pH 5.0. 4011300.
Dissolve 2.72 g of potassium dihydrogen phosphate R in
800 ml of water R. Adjust the pH (2.2.3) with 1 M potassium Mix 63.2 ml of a 71.5 g/l solution of disodium hydrogen
hydroxide and dilute to 1000 ml with water R.
Buffer solution pH 5.2. 4001700.
Dissolve 1.02 g of potassium hydrogen phthalate R in
30.0 ml of 0.1 M sodium hydroxide. Dilute to 100.0 ml with
water R.
General Notices (1) apply to all monographs and other texts
4.1.3. Buffer solutions
0.067 M Phosphate buffer solution pH 5.4. 4012000.
Mix appropriate volumes of a 23.99 g/l solution of disodium
hydrogen phosphate R with a 9.12 g/l solution of sodium
dihydrogen phosphate monohydrate R to obtain pH 5.4
(2.2.3).
Acetate-edetate buffer solution pH 5.5. 4001900.
Dissolve 250 g of ammonium acetate R and 15 g sodium
edetate R in 400 ml of water R and add 125 ml of glacial
acetic acid R.
Buffer solution pH 5.5. 4001800.
heating to 35 °C if necessary. After cooling, slowly add 10 ml
of anhydrous acetic acid R. Shake and dilute to 100.0 ml
with water R.
Phosphate buffer solution pH 5.5. 4002000.
phosphate R in water R and dilute to 1000.0 ml with the
same solvent.
Solution II. Dissolve 35.81 g of disodium hydrogen
phosphate R in water R and dilute to 1000.0 ml with the
same solvent.
Mix 96.4 ml of solution I and 3.6 ml of solution II.
Phosphate-citrate buffer solution pH 5.5. 4008700.
Mix 56.85 ml of a 28.4 g/l solution of anhydrous disodium
hydrogen phosphate R and 43.15 ml of a 21 g/l solution
of citric acid R.
Phosphate buffer solution pH 5.6. 4011200.
Solution I. Dissolve 0.908 g of potassium dihydrogen
phosphate R in water R and dilute to 100.0 ml with the
same solvent.
Solution II. Dissolve 1.161 g of dipotassium hydrogen
phosphate R in water R and dilute to 100.0 ml with the
same solvent.
Mix 94.4 ml of solution I and 5.6 ml of solution II. If
necessary, adjust to pH 5.6 (2.2.3) using solution I or
solution II.
Phosphate buffer solution pH 5.8. 4002100.
Dissolve 1.19 g of disodium hydrogen phosphate dihydrate R
and 8.25 g of potassium dihydrogen phosphate R in water R
and dilute to 1000.0 ml with the same solvent.
Acetate buffer solution pH 6.0. 4002200.
Dissolve 100 g of ammonium acetate R in 300 ml of water R,
add 4.1 ml of glacial acetic acid R, adjust the pH (2.2.3) if
necessary using ammonia R or acetic acid R and dilute to
500.0 ml with water R.
Diethylammonium phosphate buffer solution pH 6.0.
4002300.
Dilute 68 ml of phosphoric acid R to 500 ml with water R.
To 25 ml of this solution add 450 ml of water R and 6 ml of
diethylamine R, adjust to pH 6 ± 0.05 (2.2.3), if necessary,
using diethylamine R or phosphoric acid R and dilute to
500.0 ml with water R.
Phosphate buffer solution pH 6.0. 4002400.
phosphate R and 36.8 ml of a 21 g/l solution of citric acid R.
Phosphate buffer solution pH 6.0 R1. 4002500.
Dissolve 6.8 g of sodium dihydrogen phosphate R in water R
and dilute to 1000.0 ml with water R. Adjust the pH (2.2.3)
with strong sodium hydroxide solution R.
4933
4.1.3. Buffer solutions EUROPEAN PHARMACOPOEIA 5.7

Phosphate buffer solution pH 6.0 R2. 4002600.
To 250.0 ml of 0.2 M potassium dihydrogen phosphate R
add 28.5 ml of 0.2 M sodium hydroxide and dilute to
1000.0 ml with water R.
Phosphate buffer solution pH 6.4. 4002800.
Dissolve 2.5 g of disodium hydrogen phosphate R, 2.5 g
of sodium dihydrogen phosphate R and 8.2 g of sodium
chloride R in 950 ml of water R. Adjust the pH (2.2.3) of
the solution to 6.4 with 1 M sodium hydroxide or 1 M
hydrochloric acid, if necessary. Dilute to 1000.0 ml with
water R.
0.5 M Phthalate buffer solution pH 6.4. 4009200.
Dissolve 100 g of potassium hydrogen phthalate R in
water R and dilute to 1000.0 ml with the same solvent.
Adjust the pH (2.2.3) if necessary, using strong sodium
hydroxide solution R.
Buffer solution pH 6.5. 4002900.
Dissolve 60.5 g of disodium hydrogen phosphate R and
46 g of potassium dihydrogen phosphate R in water R. Add
100 ml of 0.02 M sodium edetate and 20 mg of mercuric
chloride R and dilute to 1000.0 ml with water R.
Imidazole buffer solution pH 6.5. 4003000.
Dissolve 6.81 g of imidazole R, 1.23 g of magnesium
sulphate R and 0.73 g of calcium sulphate R in 752 ml of
0.1 M hydrochloric acid. Adjust the pH (2.2.3) if necessary
and dilute to 1000.0 ml with water R.
0.1 M phosphate buffer solution pH 6.5. 4010800.
Dissolve 13.80 g of sodium dihydrogen phosphate
monohydrate R in 900 ml of distilled water R. Adjust the
pH (2.2.3) using a 400 g/l solution of sodium hydroxide R.
Dilute to 1000 ml with distilled water R.
Phosphate buffer solution pH 6.5. 4012800.
Dissolve 2.75 g of sodium dihydrogen phosphate R and
4.5 g of sodium chloride R in 500 ml of water R. Adjust thesame solvent.
pH (2.2.3) with phosphate buffer solution pH 8.5 R.
Buffer solution pH 6.6. 4003100.
To 250.0 ml of 0.2 M potassium dihydrogen phosphate R
add 89.0 ml of 0.2 M sodium hydroxide. Dilute to 1000.0 ml Mix 38.9 ml of solution I and 61.1 ml of solution II. Adjust
with water R.
Buffer solution pH 7.0. 4003500.
To 1000 ml of a solution containing 18 g/l of disodium
hydrogen phosphate R and 23 g/l of sodium chloride R add
sufficient (about 280 ml) of a solution containing 7.8 g/l
of sodium dihydrogen phosphate R and 23 g/l of sodium
chloride R to adjust the pH (2.2.3). Dissolve in the solution
sufficient sodium azide R to give a 0.2 g/l solution.
Maleate buffer solution pH 7.0. 4003600.
Dissolve 10.0 g of sodium chloride R, 6.06 g of
tris(hydroxymethyl)aminomethane R and 4.90 g of maleic
anhydride R in 900 ml of water R. Adjust the pH (2.2.3)
using a 170 g/l solution of sodium hydroxide R. Dilute to
1000.0 ml with water R.
Storage : at 2 °C to 8 °C ; use within 3 days.
0.025 M Phosphate buffer solution pH 7.0. 4009400.
Mix 1 volume of 0.063 M phosphate buffer solution pH 7.0 R
with 1.5 volumes of water R.
0.03 M Phosphate buffer solution pH 7.0. 4010300.
Dissolve 5.2 g of dipotassium hydrogen phosphate R in
900 ml of water for chromatography R. Adjust the solution
to pH 7.0 ± 0.1 using phosphoric acid R and dilute to
1000 ml with water for chromatography R.
0.05 M Phosphate buffer solution pH 7.0. 4012400.
Mix 34 ml of water R and 100 ml of 0.067 M phosphate
buffer solution pH 7.0 R.
0.063 M Phosphate buffer solution pH 7.0. 4009500.
Dissolve 5.18 g of anhydrous disodium hydrogen
phosphate R and 3.65 g of sodium dihydrogen phosphate
monohydrate R in 950 ml of water R and adjust the pH
(2.2.3) with phosphoric acid R ; dilute to 1000.0 ml with
water R.
0.067 M Phosphate buffer solution pH 7.0. 4003800.
Solution I. Dissolve 0.908 g of potassium dihydrogen
phosphate R in water R and dilute to 100.0 ml with the
Solution II. Dissolve 2.38 g of disodium hydrogen
phosphate R in water R and dilute to 100.0 ml with the
same solvent.
the pH (2.2.3) if necessary.

Phosphate buffered saline pH 6.8. 4003200.
Dissolve 1.0 g of potassium dihydrogen phosphate R, 2.0 g
of dipotassium hydrogen phosphate R and 8.5 g of sodium
chloride R in 900 ml of water R, adjust the pH (2.2.3) if
necessary and dilute to 1000.0 ml with the same solvent.
0.1 M Phosphate buffer solution pH 7.0. 4008200.
Dissolve 1.361 g of potassium dihydrogen phosphate R in
water R and dilute to 100.0 ml with the same solvent. Adjust
the pH (2.2.3) using a 35 g/l solution of disodium hydrogen
phosphate R.

Phosphate buffer solution pH 6.8. 4003300. Phosphate buffer solution pH 7.0. 4003700.
Mix 77.3 ml of a 71.5 g/l solution of disodium hydrogen Mix 82.4 ml of a 71.5 g/l solution of disodium hydrogen
phosphate R with 22.7 ml of a 21 g/l solution of citric acid R. phosphate R with 17.6 ml of a 21 g/l solution of citric acid R.
Phosphate buffer solution pH 7.0 R1. 4003900.
Phosphate buffer solution pH 6.8 R1. 4003400.
Mix 250.0 ml of 0.2 M potassium dihydrogen phosphate R
To 51.0 ml of a 27.2 g/l solution of potassium dihydrogen and 148.2 ml of a 8 g/l solution of sodium hydroxide R,
phosphate R add 49.0 ml of a 71.6 g/l solution of disodium adjust the pH (2.2.3) if necessary. Dilute to 1000.0 ml with
hydrogen phosphate R. Adjust the pH (2.2.3) if necessary. water R.
Storage : at 2 °C to 8 °C.
Phosphate buffer solution pH 7.0 R2. 4004000.
1 M tris-hydrochloride buffer solution pH 6.8. 4009300. Mix 50.0 ml of a 136 g/l solution of potassium dihydrogen
Dissolve 60.6 g of tris(hydroxymethyl)aminomethane R in phosphate R with 29.5 ml of 1 M sodium hydroxide and
400 ml of water R. Adjust the pH (2.2.3) with hydrochloric dilute to 100.0 ml with water R. Adjust the pH (2.2.3) to
acid R and dilute to 500.0 ml with water R. 7.0 ± 0.1.

4934 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 5.7 4.1.3. Buffer solutions

Phosphate buffer solution pH 7.0 R3. 4008600. Buffer solution pH 7.4. 4004600.
Dissolve 5 g of potassium dihydrogen phosphate R and 11 g Dissolve 0.6 g of potassium dihydrogen phosphate R, 6.4 g
of dipotassium hydrogen phosphate R in 900 ml of water R. of disodium hydrogen phosphate R and 5.85 g of sodium
Adjust to pH 7.0 (2.2.3) with dilute phosphoric acid R or chloride R in water R, and dilute to 1000.0 ml with the same
dilute sodium hydroxide solution R. Dilute to 1000 ml with solvent. Adjust the pH (2.2.3) if necessary.
water R and mix.
Phosphate buffered saline pH 7.4. 4005000.
Phosphate buffer solution pH 7.0 R4. 4010200. Dissolve 2.38 g of disodium hydrogen phosphate R, 0.19 g
Dissolve 28.4 g of anhydrous disodium hydrogen of potassium dihydrogen phosphate R and 8.0 g of sodium
phosphate R and 18.2 g of potassium dihydrogen chloride R in water. Dilute to 1000.0 ml with the same
phosphate R in water R and dilute to 500 ml with the same solvent. Adjust the pH (2.2.3) if necessary.
solvent.
Phosphate buffer solution pH 7.4. 4004800.
Phosphate buffer solution pH 7.0 R5. 4011400. Add 250.0 ml of 0.2 M potassium dihydrogen phosphate R
Dissolve 28.4 g of anhydrous disodium hydrogen to 393.4 ml of 0.1 M sodium hydroxide.
phosphate R in 800 ml of water R. Adjust the pH (2.2.3)
using a 30 per cent m/m solution of phosphoric acid R and Tris(hydroxymethyl)aminomethane buffer solution pH 7.4.
dilute to 1000 ml with water R. 4012100.
Dissolve 30.3 g of tris(hydroxymethyl)aminomethane R
Tetrabutylammonium buffer solution pH 7.0. 4010900. in approximately 200 ml of water R. Add 183 ml of 1 M
Dissolve 6.16 g of ammonium acetate R in a mixture of 15 ml hydrochloric acid. Dilute to 500.0 ml with water R. Note :
of tetrabutylammonium hydroxide solution (400 g/l) R and the pH is 7.7-7.8 at room temperature and 7.4 at 37 °C. This
185 ml of water R. Adjust the pH (2.2.3) with nitric acid R. solution is stable for several months at 4 °C.

Buffered salt solution pH 7.2. 4004300. Tris(hydroxymethyl)aminomethane sodium chloride buffer

Dissolve in water R 8.0 g of sodium chloride R, 0.2 g
of potassium chloride R, 0.1 g of anhydrous calcium
chloride R, 0.1 g of magnesium chloride R, 3.18 g of
disodium hydrogen phosphate R and 0.2 g of potassium
dihydrogen phosphate R and dilute to 1000.0 ml with
water R.
solution pH 7.4. 4004900.
Dissolve 6.08 g of tris(hydroxymethyl)aminomethane R,
8.77 g of sodium chloride R in 500 ml of distilled water R.
Add 10.0 g of bovine albumin R. Adjust the pH (2.2.3) using
hydrochloric acid R. Dilute to 1000.0 ml with distilled
water R.

Buffer solution pH 7.2. 4004100. Tris(hydroxymethyl)aminomethane sodium chloride buffer

To 250.0 ml of 0.2 M potassium dihydrogen phosphate R solution pH 7.4 R1. 4012200.
add 175.0 ml of 0.2 M sodium hydroxide. Dilute to 1000.0 ml Dissolve 0.1 g of bovine albumin R in a mixture containing
with water R. Adjust the pH (2.2.3) if necessary. 2 ml of tris(hydroxymethyl)aminomethane buffer solution
pH 7.4 R and 50 ml of a 5.84 mg/ml solution of sodium
Phosphate-albumin buffered saline pH 7.2. 4004400. chloride R. Dilute to 100.0 ml with water R.
Dissolve 10.75 g of disodium hydrogen phosphate R, 7.6 g
of sodium chloride R and 10 g of bovine albumin R in Tris-sodium acetate buffer solution pH 7.4. 4012900.
water R and dilute to 1000.0 ml with the same solvent. Dissolve 6.3 g of tris(hydroxymethyl)aminomethane R and
Immediately before use adjust the pH (2.2.3) using dilute 4.9 g of anhydrous sodium acetate R in 900 ml of water R.
sodium hydroxide solution R or dilute phosphoric acid R. Adjust to pH 7.4 (2.2.3) with sulphuric acid R and dilute to
1000 ml with water R.
Phosphate-albumin buffered saline pH 7.2 R1. 4009600.
Dissolve 10.75 g of disodium hydrogen phosphate R, 7.6 g Tris-sodium acetate-sodium chloride buffer solution
of sodium chloride R and 1 g of bovine albumin R in pH 7.4. 4013000.
water R and dilute to 1000.0 ml with the same solvent. Dissolve 30.0 g of tris(hydroxymethyl)aminomethane R,
Immediately before use adjust the pH (2.2.3) using dilute 14.5 g of anhydrous sodium acetate R and 14.6 g of sodium
sodium hydroxide solution R or dilute phosphoric acid R. chloride R in 900 ml of water R. Add 0.50 g of bovine
albumin R. Adjust to pH 7.4 (2.2.3) with sulphuric acid R
Phosphate buffer solution pH 7.2. 4004200. and dilute to 1000 ml with water R.
Mix 87.0 ml of a 71.5 g/l solution of disodium hydrogen
phosphate R with 13.0 ml of a 21 g/l solution of citric acid R. Borate buffer solution pH 7.5. 4005200.
Dissolve 2.5 g of sodium chloride R, 2.85 g of disodium
Imidazole buffer solution pH 7.3. 4004500. tetraborate R and 10.5 g of boric acid R in water R and
Dissolve 3.4 g of imidazole R and 5.8 g of sodium chloride R dilute to 1000.0 ml with the same solvent. Adjust the pH
in water R, add 18.6 ml of 1 M hydrochloric acid and dilute (2.2.3) if necessary.
to 1000.0 ml with water R. Adjust the pH (2.2.3) if necessary. Storage : at 2 °C to 8 °C.

Barbital buffer solution pH 7.4. 4004700.
Mix 50 ml of a solution in water R containing 19.44 g/l of
sodium acetate R and 29.46 g/l of barbital sodium R with
50.5 ml of 0.1 M hydrochloric acid, add 20 ml of an 85 g/l
of sodium chloride R and dilute to 250 ml with water R.
Buffer (HEPES) solution pH 7.5. 4009700.
Dissolve 2.38 g of 2-[4-(2-hydroxyethyl)piperazin-1-
yl]ethanesulphonic acid R in about 90 ml of water R. Adjust
the pH to 7.5 with sodium hydroxide solution R. Dilute to
100 ml with water R.

General Notices (1) apply to all monographs and other texts 4935
4.1.3. Buffer solutions EUROPEAN PHARMACOPOEIA 5.7

0.2 M Phosphate buffer solution pH 7.5. 4005400.
Dissolve 27.22 g of potassium dihydrogen phosphate R in
930 ml of water R, adjust to pH 7.5 (2.2.3) with a 300 g/l
solution of potassium hydroxide R and dilute to 1000.0 ml
with water R.
1 M Tris-hydrochloride buffer solution pH 8.0. 4012700.
Dissolve 121.1 g of tris(hydroxymethyl)aminomethane R
and 1.47 g of calcium chloride R in 900 ml of water R.
Adjust the pH (2.2.3) with hydrochloric acid R and dilute
to 1000.0 ml with water R.

0.33 M Phosphate buffer solution pH 7.5. 4005300. Tris-hydrochloride buffer solution pH 8.0. 4012300.
Solution I. Dissolve 119.31 g of disodium hydrogen Dissolve 1.21 g of tris(hydroxymethyl)aminomethane R and
phosphate R in water R and dilute to 1000.0 ml with the 29.4 mg of calcium chloride R in water R. Adjust the pH
same solvent. (2.2.3) with 1 M hydrochloric acid and dilute to 100.0 ml
with water R.
Solution II. Dissolve 45.36 g of potassium dihydrogen
phosphate R in water R and dilute to 1000.0 ml with the Tris-sodium acetate buffer solution pH 8.0. 4013100.
same solvent. Dissolve 6.3 g of tris(hydroxymethyl)aminomethane R and
Mix 85 ml of solution I and 15 ml of solution II. Adjust the 4.9 g of anhydrous sodium acetate R in 900 ml of water R.
pH (2.2.3) if necessary. Adjust to pH 8.0 (2.2.3) with sulphuric acid R and dilute to
1000 ml with water R.
0.05 M Tris-hydrochloride buffer solution pH 7.5.
4005600. Tris-sodium acetate-sodium chloride buffer solution
pH 8.0. 4013200.
Dissolve 6.057 g of tris(hydroxymethyl)aminomethane R in
Dissolve 30.0 g of tris(hydroxymethyl)aminomethane R,
water R and adjust the pH (2.2.3) with hydrochloric acid R.
Dilute to 1000.0 ml with water R. 14.5 g of anhydrous sodium acetate R and 14.6 g of sodium
chloride R in 900 ml of water R. Add 0.50 g of bovine
Tris(hydroxymethyl)aminomethane buffer solution pH 7.5. albumin R. Adjust to pH 8.0 (2.2.3) with sulphuric acid R
4005500. and dilute to 1000 ml with water R.
Dissolve 7.27 g of tris(hydroxymethyl)aminomethane R and Tris(hydroxymethyl)aminomethane buffer solution pH 8.1.
5.27 g of sodium chloride R in water R, and adjust the pH 4006200.
(2.2.3) if necessary. Dilute to 1000.0 ml with water R.
Dissolve 0.294 g of calcium chloride R in 40 ml of
Sodium citrate buffer solution pH 7.8 (0.034 M sodium tris(hydroxymethyl)aminomethane solution R and adjust
citrate, 0.101 M sodium chloride). 4009800. the pH (2.2.3) with 1 M hydrochloric acid. Dilute to 100.0 ml
with water R.
Dissolve 10.0 g of sodium citrate R and 5.90 g of sodium
chloride R in 900 ml of water R. Adjust the pH (2.2.3) by Tris-glycine buffer solution pH 8.3. 4006300.
addition of hydrochloric acid R and dilute to 1000 ml with Dissolve 6.0 g of tris(hydroxymethyl)aminomethane R
water R. and 28.8 g of glycine R in water R and dilute to 1000.0 ml
with the same solvent. Dilute 1 volume to 10 volumes with
0.0015 M Borate buffer solution pH 8.0. 4006000. water R immediately before use.
Dissolve 0.572 g of disodium tetraborate R and 2.94 g of
calcium chloride R in 800 ml of water R. Adjust the pH Tris-hydrochloride buffer solution pH 8.3. 4011800.
(2.2.3) with 1 M hydrochloric acid. Dilute to 1000.0 ml with Dissolve 9.0 g of tris(hydroxymethyl)aminomethane R
water R. in 2.9 litres of water R. Adjust the pH (2.2.3) with 1 M
hydrochloric acid. Adjust the volume to 3 litres with
Buffer solution pH 8.0. 4005900. water R.
To 50.0 ml of 0.2 M potassium dihydrogen phosphate R
add 46.8 ml of 0.2 M sodium hydroxide. Dilute to 200.0 ml Barbital buffer solution pH 8.4. 4006400.
with water R. Dissolve 8.25 g of barbital sodium R in water R and dilute
to 1000.0 ml with the same solvent.
Buffer solution pH 8.0 R1. 4010400.
Tris-EDTA BSA buffer solution pH 8.4. 4006500.
Dissolve 20 g of dipotassium hydrogen phosphate R in
900 ml of water R. Adjust the pH (2.2.3) with phosphoric Dissolve 6.1 g of tris(hydroxymethyl)aminomethane R, 2.8 g
acid R. Dilute to 1000 ml with water R. of sodium edetate R, 10.2 g of sodium chloride R and 10 g of
bovine albumin R in water R, adjust to pH 8.4 (2.2.3) using
0.02 M Phosphate buffer solution pH 8.0. 4006100. 1 M hydrochloric acid and dilute to 1000.0 ml with water R.
To 50.0 ml of 0.2 M potassium dihydrogen phosphate R Tris(hydroxymethyl)aminomethane-EDTA buffer solution
add 46.8 ml of 0.2 M sodium hydroxide. Dilute to 500.0 ml pH 8.4. 4006600.
with water R.
Dissolve 5.12 g of sodium chloride R, 3.03 g of
0.1 M Phosphate buffer solution pH 8.0. 4008400. tris(hydroxymethyl)aminomethane R and 1.40 g of sodium
edetate R in 250 ml of distilled water R. Adjust the pH
Dissolve 0.523 g of potassium dihydrogen phosphate R and (2.2.3) to 8.4 using hydrochloric acid R. Dilute to 500.0 ml
16.73 g of dipotassium hydrogen phosphate R in water R with distilled water R.
and dilute to 1000.0 ml with the same solvent.
Phosphate buffer solution pH 8.5. 4013300.
1 M Phosphate buffer solution pH 8.0. 4007800. Dissolve 3.5 g of dipotassium hydrogen phosphate R and
Dissolve 136.1 g of potassium dihydrogen phosphate R in 4.5 g of sodium chloride R in 500 ml of water R. Adjust
water R, adjust the pH (2.2.3) with 1 M sodium hydroxide. the pH (2.2.3) with a mixture of equal volumes of dilute
Dilute to 1000.0 ml with water R. phosphoric acid R and water R.

4936 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 5.7
Tris acetate buffer solution pH 8.5. 4006700.
Dissolve 0.294 g of calcium chloride R and 12.11 g of
tris(hydroxymethyl)aminomethane R in water R. Adjust
the pH (2.2.3) with acetic acid R. Dilute to 1000.0 ml with
water R.
Barbital buffer solution pH 8.6 R1. 4006900.
Dissolve in water R 1.38 g of barbital R, 8.76 g of barbital
sodium R and 0.38 g of calcium lactate R and dilute to
1000.0 ml with the same solvent.
1.5 M tris-hydrochloride buffer solution pH 8.8. 4009900.
Dissolve 90.8 g of tris(hydroxymethyl)aminomethane R in
400 ml of water R. Adjust the pH (2.2.3) with hydrochloric
acid R and dilute to 500.0 ml with water R.
Buffer (phosphate) solution pH 9.0. 4008300.
Dissolve 1.74 g of potassium dihydrogen phosphate R in
80 ml of water R, adjust the pH (2.2.3) with 1 M potassium
hydroxide and dilute to 100.0 ml with water R.
Buffer solution pH 9.0. 4007000.
Solution I. Dissolve 6.18 g of boric acid R in 0.1 M potassium in distilled water R and dilute to 1000.0 ml with the same
chloride R and dilute to 1000.0 ml with the same solvent.
Solution II. 0.1 M sodium hydroxide.
Mix 1000.0 ml of solution I and 420.0 ml of solution II.
Buffer solution pH 9.0 R1. 4007100.
Dissolve 6.20 g of boric acid R in 500 ml of water R and
adjust the pH (2.2.3) with 1 M sodium hydroxide (about
41.5 ml). Dilute to 1000.0 ml with water R.
Ammonium chloride buffer solution pH 9.5. 4007200.
Dissolve 33.5 g of ammonium chloride R in 150 ml of
water R, add 42.0 ml of concentrated ammonia R and dilute
to 250.0 ml with water R.
Storage : in a polyethylene container.
Ammonium chloride buffer solution pH 10.0. 4007300.
Dissolve 5.4 g of ammonium chloride R in 20 ml of water R,
add 35.0 ml of ammonia R and dilute to 100.0 ml with
water R.
Diethanolamine buffer solution pH 10.0. 4007500.
Dissolve 96.4 g of diethanolamine R in water R and dilute
to 400 ml with the same solvent. Add 0.5 ml of an 186 g/l
solution of magnesium chloride R and adjust the pH (2.2.3)
with 1 M hydrochloric acid. Dilute to 500.0 ml with water R.
0.1 M Ammonium carbonate buffer solution pH 10.3.
4011900.
Dissolve 7.91 g of ammonium carbonate R in 800 ml of
water R. Adjust the pH (2.2.3) with dilute sodium hydroxide
solution R. Dilute to 1000.0 ml with water R.
Ammonium chloride buffer solution pH 10.4. 4011000.
Dissolve 70 g of ammonium chloride R in 200 ml of water R,
add 330 ml of concentrated ammonia R and dilute to
1000.0 ml with water R. If necessary, adjust to pH 10.4 with
ammonia R.
Borate buffer solution pH 10.4. 4011100.
Dissolve 24.64 g of boric acid R in 900 ml of distilled
water R. Adjust the pH (2.2.3) using a 400 g/l solution
of sodium hydroxide R. Dilute to 1000 ml with distilled
water R.
General Notices (1) apply to all monographs and other texts
4.2.1. Primary standards for volumetric solutions
Ammonium chloride buffer solution pH 10.7. 4013400.
Dissolve 67.5 g of ammonium chloride R in water R, add
570 ml of concentrated ammonia R and dilute to 1000.0 ml
with water R.
Buffer solution pH 10.9. 4007600.
Dissolve 6.75 g of ammonium chloride R in ammonia R
and dilute to 100.0 ml with the same solvent.
Total-ionic-strength-adjustment buffer. 4007700.
Dissolve 58.5 g of sodium chloride R, 57.0 ml of glacial
acetic acid R, 61.5 g of sodium acetate R and 5.0 g of
cyclohexylene-dinitrilotetra-acetic acid R in water R and
dilute to 500.0 ml with the same solvent. Adjust to pH 5.0
to 5.5 with a 335 g/l solution of sodium hydroxide R and
dilute to 1000.0 ml with distilled water R.
Total-ionic-strength-adjustment buffer R1. 4008800.
Solution (a). Dissolve 210 g of citric acid R in 400 ml of
distilled water R. Adjust to pH 7.0 (2.2.3) with concentrated
ammonia R. Dilute to 1000.0 ml with distilled water R.
Solution (b). Dissolve 132 g of ammonium phosphate R
solvent.
Solution (c). To a suspension of 292 g of
(ethylenedinitrilo)tetra-acetic acid R in about 500 ml
of distilled water R, add about 200 ml of concentrated
ammonia R to dissolve. Adjust the pH to 6 to 7 (2.2.3) with
concentrated ammonia R. Dilute to 1000.0 ml with distilled
water R.
Mix equal volumes of solution (a), (b), and (c) and adjust to
pH 7.5 with concentrated ammonia R.
4.2. VOLUMETRIC ANALYSIS
04/2007:40201
4.2.1. PRIMARY STANDARDS FOR
VOLUMETRIC SOLUTIONS
Primary standards for volumetric solutions are indicated
by the suffix RV. Primary standards of suitable quality may
be obtained from commercial sources or prepared by the
following methods.
Arsenious trioxide. As2O3. (Mr 197.8). 2000100. [1327-53-3].
Sublime arsenious trioxide R in a suitable apparatus.
Storage : over anhydrous silica gel R.
Benzoic acid. C7H6O2. (Mr 122.1). 2000200. [65-85-0].
Sublime benzoic acid R in a suitable apparatus.
Potassium bromate. KBrO3. (Mr 167.0). 2000300.
[7758-01-2].
Crystallise potassium bromate R from boiling water R.
Collect the crystals and dry to constant mass at 180 °C.
Potassium hydrogen phthalate. C8H5KO4. (Mr 204.2).
2000400. [877-24-7].
Recrystallise potassium hydrogen phthalate R from boiling
water R, collect the crystals at a temperature above 35 °C
and dry to constant mass at 110 °C.
4937